Paediatric high grade gliomas (pHGG) are aggressive and difficult to treat central nervous system tumours. Current treatment incorporates surgery – where possible, followed by combined radio- and chemotherapy. Treatment response in these tumours is very poor, with significant resistance and severe side effects. The mean survival is <9 to 12 months after diagnosis, highlighting an urgent need to identify new treatments.
Alison’s project outline is to understand which genes paediatric high grade glioma cells rely on to survive, and to identify chemotherapeutics which interact with these genes. We believe targeting these altered genes will allow us to select chemotherapeutic drugs which target only cancerous cells, and not healthy brain cells, as this could help to improve efficacy and reduce side effects.
Once she has identified paediatric high grade glioma survival genes, she will select chemotherapeutic drugs which interact with and prevent gene function. These chemotherapeutic drugs may be novel, or repurposed drugs which are already used in the clinic to treat disease.
A 3D Tumour spheroid model for more biologically relevant drug testing.
We are using this 3D tumour cell model along with fluorescent stains to study the effect of chemotherapeutic drugs on cancer cells. Cells grown in 3D spheroids more closely resemble the biology of a tumour than conventional 2D cell culture.
Here, paediatric high grade glioma spheroids (KNS42) were stained with 3 dyes: Calcein AM(Staining Live Cells, Green), Hoechst 33342 (Staining DNA in all cells, Blue) and Propidium Iodide (Staining only DNA of Dead Cells, Red). The final shot is a composite of all three channels overlaid.
Using this model we can quickly test large numbers of chemotherapeutic drugs, determining how many live and dead cells are in the spheroid. This allows us to rapidly identify effective chemotherapeutic drugs to study in more detail.
Alison will use the above 3D tumour spheroid model to test the selected chemotherapeutics. Chemotherapeutic drugs which are effective in killing high grade glioma cells in the 3D spheroid model will be studied in more detail. The model is a good predictor for how chemotherapeutics might behave in a tumour, an important early step towards new therapies reaching the clinic.